JP7623666B2 - Ultrasound Sensitizers - Google Patents
Ultrasound Sensitizers Download PDFInfo
- Publication number
- JP7623666B2 JP7623666B2 JP2020073287A JP2020073287A JP7623666B2 JP 7623666 B2 JP7623666 B2 JP 7623666B2 JP 2020073287 A JP2020073287 A JP 2020073287A JP 2020073287 A JP2020073287 A JP 2020073287A JP 7623666 B2 JP7623666 B2 JP 7623666B2
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- Prior art keywords
- ethylene glycol
- derivative
- electron
- phenylchlorin
- withdrawing group
- Prior art date
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- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Description
本発明は、新規な電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体あるいはその薬理学的に許容される塩、それらを有効成分とする超音波化学療法（ＳＤＴ：Sonodynamic Therapy）用として使用する癌疾患治療剤、皮膚疾患治療剤、並びに抗菌剤に関する。
また、これらをＣｕやＭｎ金属錯体化することにより、更に水溶性を増大させた新規な電子求引基担持フェニルクロリン金属錯体誘導体或いはエチレングリコール金属錯体誘導体又はその薬理学的に許容される塩に関する。
The present invention relates to a novel phenylchlorin derivative or ethylene glycol derivative bearing an electron-withdrawing group, or a pharmacologically acceptable salt thereof, and to a cancer treatment agent, a skin treatment agent, and an antibacterial agent containing the same as an active ingredient for use in sonodynamic therapy (SDT).
The present invention also relates to novel phenylchlorin metal complex derivatives or ethylene glycol metal complex derivatives bearing an electron-withdrawing group, or pharmacologically acceptable salts thereof, which have been complexed with Cu or Mn to further increase water solubility.
癌の新しい治療法として、超音波化学療法（ＳＤＴ：Sonodynamic Therapy）用が行われている。これはある種のポルフィリン誘導体を静脈内注射などの方法により投与して、癌（腫瘍）組織に選択的に集積させた後、超音波を照射することにより癌組織のみを選択的に破壊することにより癌細胞を消滅させる治療法であり、ポルフィリン誘導体が有する癌組織への選択的集積性と、超音波増感作用という２つの特性を利用した治療法である。 Sonodynamic Therapy (SDT) is being used as a new cancer treatment. This is a treatment in which a certain type of porphyrin derivative is administered by intravenous injection or other methods, allowing it to selectively accumulate in cancer (tumor) tissue, and then ultrasound is irradiated to selectively destroy only the cancer tissue, eliminating the cancer cells. This treatment makes use of two properties of porphyrin derivatives: their ability to selectively accumulate in cancer tissue and their ultrasound sensitization effect.
他方、光物理化学的診断・治療法（ＰＤＴ：Photodynamic Therapy）は光照射による治療法で、ポルフィリン誘導体が有する癌組織への選択的集積性と、光増感作用という２つの特性を利用した治療法である。 On the other hand, photophysicochemical diagnosis and treatment (Photodynamic Therapy) is a treatment method that uses light irradiation and takes advantage of two properties of porphyrin derivatives: their ability to selectively accumulate in cancer tissue and their photosensitizing effect.
本発明者の１人は、このＰＤＴに使用することができるポルフィリン誘導体について鋭意研究を進めてきており、これまでにＡＴＸ－Ｓ１０をはじめとする誘導体を多数提供してきている（特許文献１）。 One of the inventors has been actively researching porphyrin derivatives that can be used in PDT, and has provided a number of derivatives, including ATX-S10 (Patent Document 1).
一方、ＳＤＴ増感剤として、これまでに癌疾患用に５－アミノレブリン酸塩酸塩（５－ＡＬＡ）や、クロリンＣｅ６－Ｓｎ錯体（Ｔ－Ｃｅ６）並びにエチレングリコール担持クロリンＳｎ錯体（ＡＣＴ４２１１）が開発されている（特許文献２、非特許文献１及び２）。
On the other hand, as SDT sensitizers, 5-aminolevulinic acid hydrochloride (5-ALA), chlorin Ce 6 -Sn complex (T-Ce 6 ), and ethylene glycol-supported chlorin Sn complex (ACT4211) have been developed for cancer diseases (
近年、５－ＡＬＡを用いたＰＤＴによる癌診断・治療が有効であることが分ってきている。
また、５－ＡＬＡを用いたＳＤＴによる癌治療の試みもなされている。
しかしながら、５－ＡＬＡはプロトポルフィリンＩＸの生合成前駆体であり、プロトポルフィリンＩＸの化合物特性、すなわち新生血管集積性が低いこと、最長波長吸収端が６３０ｎｍであること並びに光毒性が存在し、超音波による活性が弱いことから良好なＳＤＴ治療剤とは言えない。
In recent years, it has become clear that PDT using 5-ALA is effective for the diagnosis and treatment of cancer.
Attempts have also been made to treat cancer through SDT using 5-ALA.
However, 5-ALA is a biosynthetic precursor of protoporphyrin IX, and due to the compound properties of protoporphyrin IX, namely, low neovascular accumulation, a longest wavelength absorption edge of 630 nm, phototoxicity, and weak activity by ultrasound, 5-ALA cannot be said to be a good therapeutic agent for SDT.
他方、Ｔ－Ｃｅ６やＡＣＴ４２１１は植物体由来のクロロフィル誘導体で、我々の長年の研究から、動物由来でなく植物由来であることからこれらの安全性について必ずしも良好とは言えなかった。すなわち、本発明者らの開発した電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体は、血液由来のプロトポルフィリンから４～５工程を経て合成して得られた誘導体である。本発明者らが創生したクロリン類は、生体内で分解してプロトポルフィリン代謝を経て通常の代謝ルートで排出されるため、特に安全であることが示唆される。 On the other hand, T-Ce 6 and ACT4211 are chlorophyll derivatives derived from plants, and our long-term research has shown that their safety is not necessarily good because they are derived from plants, not animals. That is, the electron-withdrawing group-bearing phenylchlorin derivatives or ethylene glycol derivatives developed by the present inventors are derivatives obtained by synthesis through 4 to 5 steps from blood-derived protoporphyrin. The chlorins created by the present inventors are decomposed in vivo, metabolized by protoporphyrin, and excreted via the normal metabolic route, suggesting that they are particularly safe.
本出願人らは、ＳＤＴ用の化合物として、超音波増感剤である前願物質のクロリンＭｎ金属錯体（特許文献３）を出願してきた。
ところで、これまで提案してきたクロリン物質（特許第５１７９２４５号：特許文献４、特許第５６５１４２６号：特許文献５）は、Ｍｎ錯体化することにより光毒性が避けられる化合物であった。
The applicants have filed a patent application for a precursor substance, a chlorin Mn metal complex (Patent Document 3), which is an ultrasonic sensitizer, as a compound for SDT.
Meanwhile, the chlorin substances proposed thus far (Japanese Patent No. 5,179,245:
我々は、先に前願物質（特許文献３：特開２０１８－１９９６４９号公報）を開発したが、その発明物質には分子内にヨード分子を含むことで、少なからず問題があった。ヨード分子は放射線診断剤となるメリットがあったが、一方ヨード分子はアレルギー発症のもとになると思われており、分子内にヨード分子を含まない化合物が求められていた。 We previously developed the aforementioned substance (Patent Document 3: JP 2018-199649 A), but the invented substance contained iodine molecules within its molecule, which caused some problems. Iodine molecules have the advantage of being used as a radiological diagnostic agent, but on the other hand, iodine molecules are thought to be the cause of allergies, and so there was a demand for compounds that do not contain iodine molecules within their molecules.
一方、本発明者の一人のこれまでの研究によりポルフィリン骨格に電子求引基を結合、並びにＭｎ錯体化させれば光毒性を消去できることが分かっており、その上抱水クロラールが超音波に感度良く反応することから、２個以上の塩素分子が存在すれば良いことも分かっていた。
そこで、クロル化フェニル基を結合させたポルフィリンを開発したが（非特許文献３）、水溶性が極めて乏しくＳＤＴ増感剤として利用することが出来なかった。
On the other hand, previous research by one of the present inventors has revealed that phototoxicity can be eliminated by binding an electron-withdrawing group to the porphyrin skeleton and forming a Mn complex. Furthermore, since chloral hydrate reacts sensitively to ultrasound, it was also known that the presence of two or more chlorine molecules was sufficient.
Therefore, a porphyrin having a chlorinated phenyl group bonded thereto was developed (Non-Patent Document 3), but it had very poor water solubility and could not be used as an SDT sensitizer.
そこで、電子求引基担持フェニルクロリン化合物をこれまでの研究により癌への親和性が高まることが分かっているアスパラギン酸（特許文献１）のような多価カルボン酸基や多価水酸基を持つ官能基と結合させることにより水溶性が確保出来るものと逐次研究開発を重ねた。更により水溶性を持たせるためにＭｎ、Ｃｕ及びＦｅ錯体も検討した。 Therefore, we have been conducting research and development to ensure water solubility by combining phenylchlorin compounds bearing electron-withdrawing groups with functional groups having polyvalent carboxylic acid groups or polyvalent hydroxyl groups, such as aspartic acid (Patent Document 1), which has been shown in previous research to have high affinity for cancer. We have also investigated Mn, Cu and Fe complexes to further improve water solubility.
研究の結果、２個以上のカルボン酸を持つアミノ酸や、２個以上のエチレングリコール体を結合させて水溶性と癌親和性を併せ持つ誘導体を見出した。 As a result of their research, they discovered derivatives that combine water solubility and cancer affinity by combining amino acids with two or more carboxylic acids or two or more ethylene glycol units.
新規な本電子求引基担持フェニルフェニルクロリン誘導体又はエチレングリコール誘導体にあっては、癌治療剤のみならず軟膏剤、ローション剤等のＳＤＴ用の外用剤としても感染症の治療に有効であることを確認し、本発明を完成させるに至った。 The present invention has been completed by confirming that the novel electron-withdrawing group-bearing phenylphenylchlorin derivatives or ethylene glycol derivatives are effective not only as cancer treatment agents but also as external preparations for SDT such as ointments and lotions for the treatment of infectious diseases.
また、更なる水溶性の確保に向けては、本新規クロリン誘導体をＭｎ、Ｆｅ及びＣｕ金属錯体とすることも可能である。 To further ensure water solubility, this novel chlorin derivative can also be converted into a Mn, Fe, or Cu metal complex.
したがって本発明は、超音波化学療法（ＳＤＴ：Sonodynamic Therapy）に使用し得る電子求引基担持フェニルクロリンアミノ酸誘導体又はエチレングリコール誘導体、更には、それら電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体のＭｎ、ＣｕやＦｅ錯体を有効成分とするＳＤＴ用の癌注射剤や外用剤、特に癌治療剤、皮膚疾患治療剤、並びに抗菌剤を提供することを課題とする。 The present invention therefore aims to provide phenylchlorin amino acid derivatives or ethylene glycol derivatives carrying electron-withdrawing groups that can be used in sonodynamic therapy (SDT), as well as cancer injections and topical preparations for SDT, particularly cancer treatments, skin disease treatments, and antibacterial agents, that contain Mn, Cu, or Fe complexes of these phenylchlorin derivatives or ethylene glycol derivatives carrying electron-withdrawing groups as active ingredients.
かかる課題を解決するための本発明は、その一つの基本的態様として、次式（Ｉ）： In order to solve this problem, the present invention has, as one basic aspect, the following formula (I):
［式中、
Ｘは、次式
[Wherein,
X is represented by the following formula:
（なお、上記置換基を、以下、［化２］と称する。）
又は次式
(The above substituent will be referred to as [Chemical Formula 2] below.)
Or the following formula
（なお、上記置換基を、以下、［化３］と称する。） (The above substituent will be referred to as [Chemical formula 3] below.)
（上記式中、Ｙは電子吸引基を示し、ハロゲン原子、ＮＨ２、ＯＨ、又はＯＣＨ３である）で表される基を表し、
Ｒは、－Ｎ(ＣＨ２ＣＯＯＮａ)２［これはイミノ二酢酸の残基を示す］、アスパラギン酸残基或いはグルタミン酸残基、又は－ＮＨ(ＣＨ２ＣＨ２Ｏ)ｎＯＨ（ｎは、１から３の整数を表す）を表し、
Ｍは、Ｍｎ、Ｆｅ又はＣｕを表し、
Ｚは、配位子を示し、ＯＡｃ又はＣｌである］
で示される電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体あるいはその金属錯体又はその薬理学的に許容される塩である。
(wherein Y represents an electron-withdrawing group, which is a halogen atom, NH 2 , OH, or OCH 3 ),
R represents -N( CH2COONa ) 2 [which represents a residue of iminodiacetic acid], an aspartic acid residue, a glutamic acid residue, or -NH(CH2CH2O)nOH ( n represents an integer of 1 to 3);
M represents Mn, Fe or Cu;
Z represents a ligand and is OAc or Cl.
or a metal complex thereof, or a pharmacologically acceptable salt thereof.
より具体的な一つの態様としては、本発明は、前記式（Ｉ）が、次式（Ｉ）－Ａ型： In a more specific embodiment, the present invention relates to a compound represented by formula (I)-A type:
並びに、次式（Ｉ）－Ｂ型： And the following formula (I)-B type:
（上記各式中、Ｘ、Ｒ、Ｙ、Ｍ、Ｚ及びｎは、前記定義と同一である）
で示される、電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体あるいはその金属錯体又はその薬理学的に許容される塩である。
(In the above formulas, X, R, Y, M, Z and n are the same as defined above.)
or a metal complex thereof, or a pharmacologically acceptable salt thereof.
また本発明は、別の態様として、上記の電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体あるいはその金属錯体又はその薬理学的に許容される塩を有効成分として含有する、ＳＤＴ用の癌治療剤、皮膚疾患治療用剤、または抗菌剤であり、またＭｎ錯体の場合はＭＲＩ造影剤としても利用できる。 In another aspect, the present invention is a cancer treatment agent, skin disease treatment agent, or antibacterial agent for SDT, which contains the above-mentioned electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative, or a metal complex or a pharmacologically acceptable salt thereof as an active ingredient, and in the case of a Mn complex, can also be used as an MRI contrast agent.
より具体的には、本発明は、注射剤、軟膏剤、舌下剤或いはローション剤の形態にある上記したＳＤＴ用の癌治療剤、皮膚疾患治療用剤、または抗菌剤であり、またＭＲＩ造影剤としても利用できる。 More specifically, the present invention relates to a cancer treatment agent, skin disease treatment agent, or antibacterial agent for the above-mentioned SDT in the form of an injection, ointment, sublingual agent, or lotion, and can also be used as an MRI contrast agent.
すなわち本発明は、その基本的な態様は、上記式（Ｉ）で示される電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体、特に（Ｉ）－Ａ型、並びに（Ｉ）－Ｂ型の電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体あるいはその金属錯体、又はその薬理学的に許容される塩を利用して、ＳＤＴによる癌疾患治療、皮膚疾患治療、感染症治療を行う点に特徴を有するものである。 That is, the basic aspect of the present invention is characterized in that it uses an electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative represented by the above formula (I), particularly (I)-A type and (I)-B type electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative, or a metal complex thereof, or a pharmacologically acceptable salt thereof, to treat cancer disease, skin disease, and infectious disease by SDT.
本発明により提供される電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体あるいはその金属錯体又はその薬理学的に許容される塩は、水溶性クロリン誘導体であり、これらのクロリン誘導体を含有する注射剤、液剤、軟膏剤、舌下剤、坐剤或いはローション剤等の薬剤は、患部への集積性や浸透性が良好なものであり、光毒性もなく超音波に感受性があり、癌疾患治療、皮膚疾患治療並びに感染症治療におけるＳＤＴ療法において、患者に負担を与えることがない。
また超音波を使用できることから、光とは異なり深部にも到達し、治療自体を簡便に行える利点を有しており、新しい治療システムを提供できるものである。
The electron-withdrawing group-bearing phenylchlorin derivatives or ethylene glycol derivatives, or metal complexes or pharmacologically acceptable salts thereof provided by the present invention are water-soluble chlorin derivatives, and drugs containing these chlorin derivatives, such as injections, liquids, ointments, sublingual preparations, suppositories, and lotions, have good accumulation and permeability to affected areas, are non-phototoxic, and are sensitive to ultrasound, so that they do not impose a burden on patients in SDT therapy for the treatment of cancer diseases, skin diseases, and infectious diseases.
Furthermore, since ultrasound can be used, it can reach deep areas, unlike light, and has the advantage that the treatment itself can be carried out easily, providing a new treatment system.
さらに、感染症治療のためのＳＤＴ用抗菌剤として効果的なものであり、特に本電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体あるいはその金属錯体は水溶性を確保出来るため、非水性外用剤のみならず、水性外用製剤として用いることも可能である。
また、今日の癌治療、皮膚疾患治療並びに感染症治療用いられている抗がん剤、抗菌剤、抗生物質等は、薬剤耐性が容易に生じ易いものであるが、外部エネルギーとして超音波を用いる本発明のＳＤＴ治療システムにおいては、そのような薬剤耐性の問題が発生しない点で、その利点は、極めて特異的なものである。
Furthermore, they are effective as antibacterial agents for SDT in the treatment of infectious diseases, and in particular, since the present electron-withdrawing group-bearing phenylchlorin derivatives or ethylene glycol derivatives or their metal complexes can ensure water solubility, they can be used not only as non-aqueous topical agents but also as aqueous topical preparations.
Furthermore, while the anticancer drugs, antibacterial agents, antibiotics, etc. currently used in the treatment of cancer, skin diseases, and infectious diseases are prone to develop drug resistance, the SDT treatment system of the present invention, which uses ultrasound as the external energy, has a very unique advantage in that such drug resistance problems do not occur.
なお、本化合物群も、本発明者により既に提案している物質（特許文献３）の場合と同様に、ＮａＣｌ、メントール、ＮａＨＣＯ３やＥＤＴＡ等による添加増強効果が見られるものである。 In addition, this compound group also shows an enhancing effect when NaCl, menthol, NaHCO3 , EDTA, etc. are added, as in the case of the substance already proposed by the present inventor (Patent Document 3).
本発明が提供する、式（Ｉ）で示される電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体あるいはその金属錯体又はその薬理学的に許容される塩は、具体的には式（Ｉ）－Ａ型、或いは式（Ｉ）－Ｂ型の電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体（以下、これらを併せて、単に「クロリン誘導体」という場合もある）である。
このクロリン誘導体は、これまで本発明者らが提供してきたクロリン誘導体であるＴＯＮＳ５０３および５０４‐Ｍｎ錯体（特許文献３）と同様に、癌への親和性や皮膚浸透性が高く、しかも光毒性が無く、また水溶性或いは脂溶性を持たせたことから、例えば注射剤、あるいは各種軟膏基剤中に均一に溶解・分散し、軟膏製剤自体の安定性も極めて良好なものであり、また水溶性ローション剤として製剤中に均一に溶解・分散し、製剤自体の安定性も極めて良好なものである。
The electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative represented by formula (I) or a metal complex or a pharmacologically acceptable salt thereof provided by the present invention is specifically an electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative of formula (I)-A type or formula (I)-B type (hereinafter, these may be collectively referred to simply as "chlorin derivatives").
This chlorin derivative, like the chlorin derivatives TONS503 and 504-Mn complexes (Patent Document 3) that the present inventors have previously provided, has high affinity for cancer and high skin permeability, and is non-phototoxic. In addition, because it is water-soluble or fat-soluble, it can be uniformly dissolved and dispersed in, for example, injections or various ointment bases, and the stability of the ointment formulation itself is also extremely good. Furthermore, it can be uniformly dissolved and dispersed in formulations as water-soluble lotions, and the stability of the formulation itself is also extremely good.
そのような式（Ｉ）で示されるクロリン誘導体の中でも、特に以下の式（Ｉ）で示されるＡ型、及びＢ型： Among such chlorin derivatives represented by formula (I), particularly preferred are type A and type B represented by the following formula (I):
で示されるフェニルクロリン誘導体又はエチレングリコール誘導体あるいはその金属錯体又はその薬理学的に許容される塩であって、具体的には、以下の略号で示される化合物である。 A phenylchlorin derivative or ethylene glycol derivative represented by the formula (I) or a metal complex or a pharmacologically acceptable salt thereof, specifically, a compound represented by the following abbreviations:
Ｃｏｍｐｏｕｎｄ Ａ：式（Ｉ）Ｂ型において、Ｘが［化２］であり、ＹがＣｌで、Ｒがイミノ二酢酸残基である電子求引基担持フェニルクロリンアミノ酸誘導体。 Compound A : A phenylchlorin amino acid derivative having an electron-withdrawing group, in which, in the B type of formula (I), X is [Chemical formula 2], Y is Cl, and R is an iminodiacetic acid residue.
Ｃｏｍｐｏｕｎｄ Ｂ：式（Ｉ）Ｂ型において、Ｘが［化２］であり、ＹがＣｌで、Ｒが－ＮＨ(ＣＨ２ＣＨ２Ｏ)２ＯＨである電子求引基担持フェニルクロリンエチレングリコール誘導体。 Compound B : A phenylchlorine ethylene glycol derivative bearing an electron-withdrawing group, in which in the B type of formula (I), X is [Chemical formula 2], Y is Cl, and R is -NH(CH 2 CH 2 O) 2 OH.
Ｃｏｍｐｏｕｎｄ Ｃ：式（Ｉ）Ｂ型において、Ｘが［化２］であり、ＹがＣｌで、Ｒが－ＮＨ(ＣＨ２ＣＨ２Ｏ)２ＯＨで、ＭがＭｎである電子求引基担持フェニルクロリンエチレングリコールＭｎ金属錯体。 Compound C : An electron-withdrawing group-bearing phenylchlorine ethylene glycol Mn metal complex, in which X is [Chemical formula 2], Y is Cl, R is -NH( CH2CH2O ) 2OH , and M is Mn in the formula (I) type B.
Ｃｏｍｐｏｕｎｄ Ｄ：式（Ｉ）Ｂ型において、Ｘが［化２］であり、ＹがＦで、Ｒがイミノ二酢酸残基である電子求引基担持フェニルクロリンアミノ酸誘導体。 Compound D : A phenylchlorin amino acid derivative having an electron-withdrawing group, in which, in the B type of formula (I), X is [Chemical formula 2], Y is F, and R is an iminodiacetic acid residue.
Ｃｏｍｐｏｕｎｄ Ｅ：式（Ｉ）Ｂ型において、Ｘが［化２］であり、ＹがＦで、Ｒが－ＮＨ(ＣＨ２ＣＨ２Ｏ)２ＯＨである電子求引基担持フェニルクロリンエチレングリコール誘導体。 Compound E : A phenylchlorine ethylene glycol derivative having an electron-withdrawing group, in which X is [Chemical formula 2], Y is F, and R is -NH(CH 2 CH 2 O) 2 OH in the formula (I) type B.
Ｃｏｍｐｏｕｎｄ Ｆ：式（Ｉ）Ｂ型において、Ｘが［化３］であり、Ｒが－ＮＨ(ＣＨ２ＣＨ２Ｏ)２ＯＨであるクロリンエチレングリコール誘導体。 Compound F : A chlorine ethylene glycol derivative of formula (I) type B, in which X is [Chemical formula 3] and R is --NH(CH 2 CH 2 O) 2 OH.
Ｃｏｍｐｏｕｎｄ Ｇ：式（Ｉ）Ｂ型において、Ｘが［化３］であり、Ｒが－ＮＨ(ＣＨ２ＣＨ２Ｏ)２ＯＨで、ＭがＭｎであるクロリンエチレングリコールＭｎ金属錯体。 Compound G : A chlorine ethylene glycol Mn metal complex in which, in the B type of formula (I), X is [Chemical formula 3], R is -NH(CH 2 CH 2 O) 2 OH, and M is Mn.
これらのフェニルクロリン誘導体は、Ｃｏｍｐｏｕｎｄ ＡやＤに対するＲがイミノ二酢酸残基だけでなくアスパラギン酸残基やグルタミン酸残基でも良く、多価カルボン酸を持つアミノ酸であれば良い。 In these phenylchlorin derivatives, R in Compounds A and D may be not only an iminodiacetic acid residue but also an aspartic acid residue or a glutamic acid residue, and may be any amino acid having a polycarboxylic acid.
また、Ｃｏｍｐｏｕｎｄ Ｂ、Ｃ、Ｅ、ＦやＧに対するＲが２個の連結するエチレングリコール残基だけでなく１個や３個のエチレングリコール残基でも良い。 In addition, R for Compounds B, C, E, F, and G may be not only two linked ethylene glycol residues, but also one or three linked ethylene glycol residues.
一方、ＹはＣｌだけでなく電子求引基を示す他のハロゲン化合物すなわちＢｒ、Ｉ、Ｆ原子でもよく、また同様に電子求引基を示すＮＨ２、ＯＨ、ＯＣＨ３であっても良い。
要するに、電子求引基を担持して光毒性を消去する官能基があればよく、しかも中でもフェニル基のオルト位やパラ位の配向が最も良い。しかし、これにこだわる必要もない。
On the other hand, Y may be not only Cl but also other halogen compounds exhibiting electron-withdrawing groups, that is, Br, I, F atoms, and also NH 2 , OH, OCH 3 exhibiting electron-withdrawing groups.
In short, all that is needed is a functional group that carries an electron-withdrawing group and eliminates phototoxicity, and among these, the ortho or para orientation of the phenyl group is the best. However, there is no need to be so strict about this.
金属錯体化について、これらの合成手順を代えて、最初の段階で金属錯体化しても、また合成途中の段階でも金属錯体化しても良い。
要するに、最終的に目的化合物が得られれば、合成手順のいずれの段階でも金属錯体化しても何ら問題はない。
したがって、前記特許に記載の内容は、本願明細書の一部を構成する。
Regarding the metal complexation, these synthesis procedures may be changed and the metal complexation may be carried out at the initial stage or at any stage during the synthesis.
In short, as long as the target compound is finally obtained, there is no problem in forming a metal complex at any stage of the synthesis procedure.
The contents of said patent are therefore incorporated herein by reference.
錯体化の手段としては、前記特許に記載の方法により得られた化合物等を適当な有機溶媒中に溶解後、そこにマンガンや銅の酢酸塩や塩化物を反応させることにより、フェニルクロリン誘導体又はエチレングリコール誘導体の金属錯体を調製することができる。 As a method of complex formation, the compound obtained by the method described in the above patent can be dissolved in an appropriate organic solvent, and then reacted with manganese or copper acetate or chloride to prepare a metal complex of a phenylchlorin derivative or an ethylene glycol derivative.
反応に使用する金属塩や有機溶媒は特に限定されず、反応に直接の影響を与えないものであれば、任意に選択することができる。具体的には、金属塩としては酢酸塩や塩化金属塩を挙げることができ、なかでも塩化金属塩が好ましく使用され、また有機溶媒として酢酸、ＤＭＦ等の溶媒を挙げることができ、なかでも酢酸が好ましく使用される。 There are no particular limitations on the metal salts and organic solvents used in the reaction, and any can be selected as long as they do not have a direct effect on the reaction. Specifically, examples of metal salts include acetates and metal chlorides, with metal chlorides being preferred, and examples of organic solvents include acetic acid, DMF, and the like, with acetic acid being preferred.
反応温度、反応時間も特に限定されるものではなく、４０～８０℃、好ましくは５５℃程度の加熱下に４～１０時間程度攪拌処理をすることがよい。 There are no particular limitations on the reaction temperature or reaction time, but it is recommended to heat the mixture to 40 to 80°C, preferably 55°C, and stir for 4 to 10 hours.
一方、クロリン体を電子求引基担持フェニル誘導体化又はエチレングリコール誘導体化する際には、合成手順として最初の段階でなく工程途中の段階でも当該誘導体化しても良い。
要するに、最終的に目的化合物が得られれば、合成手順のいずれの段階でも電子求引基担持フェニル誘導体化又はエチレングリコール誘導体化しても何ら問題はない。
On the other hand, when the chlorin derivative is converted into an electron-withdrawing group-bearing phenyl derivative or an ethylene glycol derivative, the derivatization may be carried out not only at the initial stage of the synthetic procedure but also at any intermediate stage in the process.
In short, as long as the target compound is finally obtained, there is no problem whether the compound is derivatized with an electron-withdrawing group-bearing phenyl derivatization or ethylene glycol derivatization at any stage of the synthetic procedure.
その幾つかの調製方法の具体的なものを以下に示す。 Specific examples of some of the preparation methods are shown below.
製造例１：式（Ｉ）Ｂ型において、Ｘが［化２］であり、Ｙがクロル、Ｒがイミノ二酢酸残基であるジクロロフェニルクロリンアミノ酸誘導体の調製（Ｃｏｍｐｏｕｎｄ Ａの調製）Production Example 1: Preparation of dichlorophenylchlorin amino acid derivatives in which X is [Chemical formula 2], Y is chlorine, and R is an iminodiacetic acid residue in the B type of formula (I) (Preparation of Compound A)
（ａ）ＣＰ－Ｐ－Ｈの調製
フォトプロトポルフィリンＩＸ ジメチルエステル（Ｐ－ＭｅＢ型）（１０ｇ）及び 2,4-dichlorophenylhydrazine HCl塩（５ｇ）をピリジン（１００ｍＬ）に溶解し１．５時間撹拌反応した。反応後、反応液を冷やした１６％酢酸水中に加えて沈殿物を濾過し、濾集物（ＣＰ－Ｐ－Ｍｅ）を水にて洗浄し、風乾した。
得られた風乾物をＤＭＦ（１５０ｍＬ）に溶解し、１．５Ｍ－ＮａＯＨ（４０ｍＬ）を加えて室温下で１．５時間撹拌し加水分解した。加水分解後、加水分解物に冷水中を加えて沈殿させ、沈殿物を濾過し、濾集物を水で数回洗浄して加水分解物を約１２ｇ（ＣＰ－Ｐ－Ｎａ）得た。得られた加水分解物を１０％クエン酸水溶液（８００ｍＬ）に懸濁させ２．５時間撹拌した。撹拌後、濾集し、濾過物を水洗後、乾燥させ、ＣＰ－Ｐ－Ｈを約１０ｇ得た。
(a) Preparation of CP-P-H Photoprotoporphyrin IX dimethyl ester (P-MeB type) (10 g) and 2,4-dichlorophenylhydrazine HCl salt (5 g) were dissolved in pyridine (100 mL) and reacted with stirring for 1.5 hours. After the reaction, the reaction solution was added to chilled 16% acetic acid water, and the precipitate was filtered. The collected matter (CP-P-Me) was washed with water and air-dried.
The air-dried product obtained was dissolved in DMF (150 mL), 1.5 M NaOH (40 mL) was added, and the mixture was stirred at room temperature for 1.5 hours to hydrolyze. After hydrolysis, the hydrolyzate was precipitated by adding cold water, and the precipitate was filtered and the filter cake was washed several times with water to obtain about 12 g of hydrolyzate (CP-P-Na). The hydrolyzate obtained was suspended in a 10% aqueous citric acid solution (800 mL) and stirred for 2.5 hours. After stirring, the mixture was filtered, and the filter cake was washed with water and dried to obtain about 10 g of CP-P-H.
ＣＰ－Ｐ－ＨのＮＭＲ分析とＩＲ分析
1H-NMR (400 MHz, CDCl3) δ 9.72 (s, 1H), 9.54 (s, 1H), 8.61 (s, 1H), 8.43 (s, 1H), 7.74-7.99 (m, 3H), 7.03 (s, 1H), 6.03-6.13 (m, 2H), 4.13-4.38 (m, 4H), 3.69 (s, 6H), 3.46 (s, 3H), 3.18-3.36 (m, 4H), 1.26 (s, 3H)
γporphyrin = 1732, 2922, 2950 cm-1, γNH=N = 1510, 1593 cm-1
NMR and IR analysis of CP-P-H
1 H-NMR (400 MHz, CDCl 3 ) δ 9.72 (s, 1H), 9.54 (s, 1H), 8.61 (s, 1H), 8.43 (s, 1H), 7.74-7.99 (m, 3H), 7.03 (s, 1H), 6.03-6.13 (m, 2H), 4.13-4.38 (m, 4H), 3.69 (s, 6H), 3.46 (s, 3H), 3.18-3.36 (m, 4H), 1.26 (s, 3H)
γ porphyrin = 1732, 2922, 2950 cm -1 , γ NH=N = 1510, 1593 cm -1
（ｂ）イミノ二酢酸 ジメチルエステルの調製
イミノ二酢酸（１０ｇ）にメタノール（５０ｍＬ）を加え、ice bath 上懸濁状態で撹拌した。ついで、塩化チオニル（１６ｍＬ）をゆっくり滴下した。すべて滴下した後、室温で１２時間反応させた。反応後、エーテルを加え沈殿物を濾過し、濾集物をエーテルでよく洗浄を行った。洗浄した固体を回収・乾燥させ、イミノ二酢酸 ジメチルエステル・塩酸塩（ＩＤＡ(ＯＭｅ)２・ＨＣｌ塩）を約１１ｇ得た。
(b) Preparation of iminodiacetic acid dimethyl ester Methanol (50 mL) was added to iminodiacetic acid (10 g) and stirred in a suspended state on an ice bath. Then, thionyl chloride (16 mL) was slowly added dropwise. After all the ingredients were added, the reaction was carried out at room temperature for 12 hours. After the reaction, ether was added, and the precipitate was filtered, and the collected matter was washed thoroughly with ether. The washed solid was collected and dried to obtain about 11 g of iminodiacetic acid dimethyl ester hydrochloride (IDA(OMe) 2.HCl salt).
ＩＤＡ（ＯＭｅ）２・ＨＣｌ塩のＮＭＲ分析
1H-NMR (500 MHz, DMSO-D6) δ 10.05 (s, 1H), 4.01 (s, 4H), 3.74 (s, 6H)
NMR analysis of IDA(OMe) 2.HCl salt
1 H-NMR (500 MHz, DMSO-D6) δ 10.05 (s, 1H), 4.01 (s, 4H), 3.74 (s, 6H)
（ｃ）目的物（Ｃｏｍｐｏｕｎｄ Ａ）の調製
上記の（ａ）で得られたＣＰ－Ｐ－Ｈ（５．０ｇ）（７８１ｇ／ｍｏｌ）及びＥＤＣ／ＨＣｌ（１０ｇ）（１５５．２５ｇ／ｍｏｌ）をＮ，Ｎ－ジメチルアセトアミド（１００ｍＬ）に溶解させた。その後、（ｂ）で得られたＩＤＡ(ＯＭｅ)２・ＨＣｌ塩（６．０ｇ）を添加し２．５時間反応させた。そして更に４．０ｇのＥＤＣ／ＨＣｌを追加添加して１．５時間反応させた。反応後、冷水中に加え沈殿物を得た。沈殿物を濾集し、水、ＥｔＯＨにて洗浄し乾燥させＣＰ－Ｐ－ＩＤＡ(ＯＭｅ)２を約５ｇ得た。
得られたＣＰ－Ｐ－ＩＤＡ(ＯＭｅ)２をＥｔＯＨ（３００ｍＬ）で懸濁させ、１．５－Ｍ－ＮａＯＨ（５０ｍＬ）を添加し１．５時間室温にて加水分解をした。得られた加水分解物に、ＥｔＯＨ／ＡｃＯＥｔ混液（１：１）（５００ｍL）を加え、沈殿物を得た。沈殿物を濾集し、ＥｔＯＨ／ＡｃＯＥｔ混液、ＥｔＯＨで洗浄後乾燥させ、目的物のＣｏｍｐｏｕｎｄ Ａを４ｇ得た。
(c) Preparation of the target product (Compound A) CP-P-H (5.0 g) (781 g/mol) obtained in (a) above and EDC/HCl (10 g) (155.25 g/mol) were dissolved in N,N-dimethylacetamide (100 mL). Then, IDA(OMe) 2.HCl salt (6.0 g) obtained in (b) was added and reacted for 2.5 hours. Then, 4.0 g of EDC/HCl was added and reacted for 1.5 hours. After the reaction, the mixture was added in cold water to obtain a precipitate. The precipitate was collected by filtration, washed with water and EtOH, and dried to obtain about 5 g of CP-P-IDA(OMe) 2 .
The obtained CP-P-IDA(OMe) 2 was suspended in EtOH (300 mL), 1.5-M-NaOH (50 mL) was added, and hydrolysis was carried out at room temperature for 1.5 hours. To the obtained hydrolyzate, EtOH/AcOEt mixed solution (1:1) (500 mL) was added to obtain a precipitate. The precipitate was collected by filtration, washed with EtOH/AcOEt mixed solution and EtOH, and then dried to obtain 4 g of the target Compound A.
Ｃｏｍｐｏｕｎｄ Ａ（Mw:1,071.78）のＮＭＲ分析とＩＲ分析
1H-NMR (400 MHz, CDCl3) δ 9.73 (s, 1H), 9.60 (s, 1H), 8.70 (s, 1H), 8.42 (s, 1H), 8.11 (br, 1H), 7.75 (d, J = 8.4 Hz, 2H), 7.37-7.43 (m, 2H), 7.18 (br, 1H), 6.16 (d, J = 18.4 Hz, 1H), 6.05 (d, J = 12.0 Hz, 1H), 4.28-4.39 (m, 2H), 4.18-4.24 (m, 2H), 3.66-3.71 (m, 8H), 3.54 (s, 3H), 3.46 (s, 3H), 3.35 (s, 3H), 3.27 (t, J = 6.8 Hz, 2H), 3.21 (t, J = 4.4 Hz, 2H), 1.44 (s, 3H)
γporphyrin = 2956 cm-1, γCOO = 1732 cm-1, γCONRR’ = 1594 cm-1,γNH=N = 1510 cm-1
これは、ＵＶスペクトル分析によってもその構造が支持された。
NMR and IR analysis of Compound A (Mw: 1,071.78)
1 H-NMR (400 MHz, CDCl 3 ) δ 9.73 (s, 1H), 9.60 (s, 1H), 8.70 (s, 1H), 8.42 (s, 1H), 8.11 (br, 1H), 7.75 (d, J = 8.4 Hz, 2H), 7.37-7.43 (m, 2H), 7.18 (br, 1H), 6.16 (d, J = 18.4 Hz, 1H), 6.05 (d, J = 12.0 Hz, 1H), 4.28-4.39 (m, 2H), 4.18-4.24 (m, 2H), 3.66-3.71 (m, 8H), 3.54 (s, 3H), 3.46 (s, 3H), 3.35 (s, 3H), 3.27 (t, J = 6.8 Hz, 2H), 3.21 (t, J = 4.4 Hz, 2H), 1.44 (s, 3H)
γ porphyrin = 2956 cm -1 , γ COO = 1732 cm -1 , γ CONRR' = 1594 cm -1 , γ NH=N = 1510 cm -1
This structure was also supported by UV spectroscopic analysis.
製造例２：式（Ｉ）Ｂ型において、Ｘが［化２］であり、Ｙがクロル、Ｒが－ＮＨＣＨ ２ ＣＨ ２ ＯＣＨ ２ ＣＨ ２ ＯＨであるジクロロフェニルクロリンエチレングリコール誘導体の調製（Ｃｏｍｐｏｕｎｄ Ｂの調製）
先の製造例１の（ａ）で得られたＣＰ－Ｐ－Ｈ（３．０ｇ）を用い、これにＥＤＣ／ＨＣｌ（６．０ｇ）を加えてＮ，Ｎ－ジメチルアセトアミドに溶解させた。その後、2-(2-aminoethoxy)ethanol（ＤＥＧＡ）（２．６ｍＬ）を添加し、１時間反応させた。さらにＥＤＣ／ＨＣｌ（１．０ｇ）を追加添加し３時間反応させた。反応後、反応物を冷水中に加え沈殿物を得た。得られた反応物を水ついでＥｔＯＨで洗浄し、デシケーターで乾燥させ目的物のＣｏｍｐｏｕｎｄ Ｂを２．８ｇ得た。
Production Example 2: Preparation of dichlorophenyl chlorine ethylene glycol derivative in which X is [Chemical formula 2], Y is chlorine, and R is -NHCH 2 CH 2 OCH 2 CH 2 OH in the B type of formula (I) (Preparation of Compound B)
CP-P-H (3.0 g) obtained in (a) of the previous Production Example 1 was used, and EDC/HCl (6.0 g) was added thereto and dissolved in N,N-dimethylacetamide. Then, 2-(2-aminoethoxy)ethanol (DEGA) (2.6 mL) was added and reacted for 1 hour. EDC/HCl (1.0 g) was further added and reacted for 3 hours. After the reaction, the reaction product was added to cold water to obtain a precipitate. The resulting reaction product was washed with water and then EtOH, and dried in a desiccator to obtain 2.8 g of the target Compound B.
Ｃｏｍｐｏｕｎｄ Ｂ（Mw:927.93）のＮＭＲ分析とＩＲ分析
1H-NMR (400 MHz, CDCl3) δ 9.73 (s, 1H), 9.53 (s, 1H), 8.52 (s, 1H), 8.42 (s, 1H), 7.93 (br, 1H), 7.60-7.77 (m, 2H), 7.39-7.44 (m, 2H), 6.98 (br, 1H), 6.11 (d, J = 17.6 Hz, 1H), 6.03 (d, J = 11.6 Hz, 1H), 4.22-4.34 (m, 4H), 4.12 (q, J = 6.9 Hz, 4H), 3.20-3.72 (m, 25H), 1.24 (s, 3H)
γOH = 3341 cm-1, γporphyrin = 1736, 2922, 2955 cm-1, γCONR = 1593 cm-1, γNH=N = 1510 cm-1,γR-O-R’ = 1115, 1239 cm-1
これは、ＵＶスペクトル分析によってもその構造が支持された。
NMR and IR analysis of Compound B (Mw: 927.93)
1 H-NMR (400 MHz, CDCl 3 ) δ 9.73 (s, 1H), 9.53 (s, 1H), 8.52 (s, 1H), 8.42 (s, 1H), 7.93 (br, 1H), 7.60-7.77 (m, 2H), 7.39-7.44 (m, 2H), 6.98 (br, 1H), 6.11 (d, J = 17.6 Hz, 1H), 6.03 (d, J = 11.6 Hz, 1H), 4.22-4.34 (m, 4H), 4.12 (q, J = 6.9 Hz, 4H), 3.20-3.72 (m, 25H), 1.24 (s, 3H)
γ OH = 3341 cm -1 , γ porphyrin = 1736, 2922, 2955 cm -1 , γ CONR = 1593 cm -1 , γ NH=N = 1510 cm -1 , γ RO-R' = 1115, 1239 cm -1
This structure was also supported by UV spectroscopic analysis.
製造例３：式（Ｉ）Ｂ型において、Ｘが［化２］であり、Ｙがクロル、Ｒが－ＮＨＣＨ ２ ＣＨ ２ ＯＣＨ ２ ＣＨ ２ ＯＨ、ＭがＭｎであるジクロロフェニルクロリンエチレングリコールＭｎ錯体の調製（Ｃｏｍｐｏｕｎｄ Ｃの調製）
先の製造例２得られたＣｏｍｐｏｕｎｄ Ｂ（１．０ｇ）を用い、これにＤＭＦ３０ｍＬを加えて溶解後、Ｍｎ(ＯＡｃ)２を加えて５時間加熱沸騰反応させて、Ｍｎ化を行った。反応後反応物を冷水中に加え沈殿物を得た。得られた反応物を水ついでエタノールで洗浄し、デシケーター内で乾燥させて目的物のＣｏｍｐｏｕｎｄ Ｃを０．８ｇ得た。
Ｃｏｍｐｏｕｎｄ Ｃの安定性をＨＰＬＣで確認し、安定であることを確認した。またＩＣＰ分析によりＭｎイオンが１００％配位されていることも確認した。
Production Example 3: Preparation of dichlorophenylchlorin ethylene glycol Mn complex in which X is [Chemical formula 2], Y is chlorine, R is --NHCH 2 CH 2 OCH 2 CH 2 OH, and M is Mn in the B type of formula (I) (preparation of Compound C)
Compound B (1.0 g) obtained in the previous Preparation Example 2 was dissolved in 30 mL of DMF, and Mn(OAc) 2 was added and reacted with heating and boiling for 5 hours to perform Mn conversion. After the reaction, the reactant was added to cold water to obtain a precipitate. The reactant was washed with water and then with ethanol, and dried in a desiccator to obtain 0.8 g of the target Compound C.
The stability of Compound C was confirmed by HPLC, and it was found to be stable. In addition, it was confirmed by ICP analysis that Mn ions were 100% coordinated.
製造例４：式（Ｉ）Ｂ型において、Ｘが［化２］であり、Ｙがフッ素、Ｒがイミノ二酢酸残基であるジフルオロフェニルクロリンアミノ酸誘導体の調製（Ｃｏｍｐｏｕｎｄ Ｄの調製）
（ａ）ＦＰ－Ｐ－Ｈの調製
フォトプロトポルフィリンＩＸ ジメチルエステル（Ｐ－ＭｅＢ型)（１０ｇ）及び 2,4-difluorophenylhydrazine HCl塩（５ｇ）をピリジン（１００ｍＬ）に溶解し１．５時間撹拌反応した。反応後、反応液を冷やした１６％酢酸水中に加えて沈殿物を濾過し、濾集物（ＦＰ－Ｐ－Ｍｅ）を水にて洗浄し、風乾しＦＰ－Ｐ－Ｈを得た。以下は製造例１と同様に処理して目的物のＣｏｍｐｏｕｎｄ Ｄを得た。
Production Example 4: Preparation of difluorophenyl chlorin amino acid derivative in which X is [Chemical formula 2], Y is fluorine, and R is iminodiacetic acid residue in the formula (I) B type (Preparation of Compound D)
(a) Preparation of FP-P-H Photoprotoporphyrin IX dimethyl ester (P-MeB type) (10 g) and 2,4-difluorophenylhydrazine HCl salt (5 g) were dissolved in pyridine (100 mL) and reacted with stirring for 1.5 hours. After the reaction, the reaction solution was added to cooled 16% acetic acid water, and the precipitate was filtered. The collected matter (FP-P-Me) was washed with water and air-dried to obtain FP-P-H. The following treatments were performed in the same manner as in Production Example 1 to obtain the target Compound D.
製造例５：式（Ｉ）Ｂ型において、Ｘが［化２］であり、Ｙがフッ素、Ｒが－ＮＨＣＨ ２ ＣＨ ２ ＯＣＨ ２ ＣＨ ２ ＯＨであるジクロロフェニルクロリンエチレングリコール誘導体の調製（Ｃｏｍｐｏｕｎｄ Ｅの調製）
先の製造例４の（ａ）で得られたＦＰ－Ｐ－Ｈ（３．０ｇ）を用い、製造例２と同様に処理して目的物のＣｏｍｐｏｕｎｄ Ｅを得た。
Production Example 5: Preparation of dichlorophenylchlorin ethylene glycol derivative in which X is [Chemical formula 2], Y is fluorine, and R is -NHCH 2 CH 2 OCH 2 CH 2 OH in the B type of formula (I) (Preparation of Compound E)
The target compound, Compound E, was obtained by treating FP-PH (3.0 g) obtained in the above Production Example 4(a) in the same manner as in Production Example 2.
製造例６：式（Ｉ）Ｂ型において、Ｘが［化３］であり、Ｒが－ＮＨＣＨ ２ ＣＨ ２ ＯＣＨ ２ ＣＨ ２ ＯＨであるクロリンエチレングリコール誘導体の調製（Ｃｏｍｐｏｕｎｄ Ｆの調製）
（ａ）ＰＧ－Ｐ－Ｍｅの調製
先の特願２０１０－２３８３５９（特許５６５１４２６号）の製造例記載の方法と同様に反応・後処理等の操作をして、ＰＧ－Ｐ－Ｍｅを得た。
（ｂ）ＰＧ－Ｐ－ＤＥＧＡの調製
先の製造例２の方法に従って、ＰＧ－Ｐ－Ｍｅを加水分解後ＰＧ－Ｐ－Ｈとし、以下製造例２と同様に操作して、Ｃｏｍｐｏｕｎｄ Ｆ（ＰＧ－Ｐ－ＤＥＧＡ）を得た。
Production Example 6: Preparation of chlorine ethylene glycol derivative in which X is [Chemical formula 3] and R is -NHCH 2 CH 2 OCH 2 CH 2 OH in the B type of formula (I) (Preparation of Compound F)
(a) Preparation of PG-P-Me PG-P-Me was obtained by carrying out the reaction, post-treatment, and other operations in the same manner as described in the production example of the above-mentioned Japanese Patent Application No. 2010-238359 (Patent No. 5651426).
(b) Preparation of PG-P-DEGA According to the method of Production Example 2, PG-P-Me was hydrolyzed to give PG-P-H, and the rest of the procedure was carried out in the same manner as in Production Example 2 to obtain Compound F (PG-P-DEGA).
製造例７：式（Ｉ）Ｂ型において、Ｘが［化３］であり、Ｒが－ＮＨＣＨ ２ ＣＨ ２ ＯＣＨ ２ ＣＨ ２ ＯＨで、ＭがＭｎであるクロリンエチレングリコールＭｎ錯体の調製（Ｃｏｍｐｏｕｎｄ Ｇの調製）
先の製造例６の方法に従って、同様に操作してＣｏｍｐｏｕｎｄ Ｆを得た後、Ｍｎ錯体化して目的とするＣｏｍｐｏｕｎｄ Ｇ（ＰＧ－Ｐ－ＤＥＧＡ Ｍｎ錯体）を得た。
Production Example 7: Preparation of chlorine ethylene glycol Mn complex in which X is [Chemical formula 3], R is -NHCH 2 CH 2 OCH 2 CH 2 OH, and M is Mn in the B type of formula (I) (preparation of Compound G)
Compound F was obtained by the same procedure as in Preparation Example 6, and then the compound was converted into a manganese complex to obtain the desired compound G (PG-P-DEGA Mn complex).
本発明においては、これらのフェニルクロリン誘導体又はエチレングリコール誘導体あるいはそれらの金属錯体は、ＳＤＴ用の癌疾患治療剤等の注射剤および皮膚疾患治療剤、或いは抗菌剤等の外用剤として処方される。
癌疾患治療剤としての注射剤は，浸透圧やｐＨ調整剤を用い、舌下剤は適当なゲル化剤等を用いて調製し、皮膚疾患治療剤、或いは抗菌剤としての外用剤は、非水性軟膏剤、水性軟膏剤、ローション剤等の剤型で処方される。
In the present invention, these phenylchlorin derivatives or ethylene glycol derivatives or metal complexes thereof are formulated as injectable preparations for SDT such as cancer disease treatments, skin disease treatments, or external preparations such as antibacterial agents.
Injectable preparations for treating cancer diseases are prepared using osmotic pressure or pH adjusters, and sublingual preparations are prepared using appropriate gelling agents, etc., and topical preparations for treating skin diseases or antibacterial agents are prescribed in the form of non-aqueous ointments, aqueous ointments, lotions, etc.
本発明において、これらの製剤に含有させる式（Ｉ）で示されるフェニルクロリン誘導体又はエチレングリコール誘導体あるいはその金属錯体の配合量は、配合された有効成分であるフェニルクロリン誘導体又はエチレングリコール誘導体あるいはその金属錯体が患部部位に到達し残留され、また経皮吸収され、疾患部位に蓄積され、超音波の照射により標的細胞、或いは細菌を死滅させるのに十分な量が配合されればよい。本発明者らの検討によれば、その配合量は、製剤重量をベースとして０．１～２０重量％であれば、十分な効果が得られることが判明した。 In the present invention, the amount of the phenylchlorin derivative or ethylene glycol derivative or metal complex thereof represented by formula (I) contained in these preparations may be an amount sufficient for the active ingredient, the phenylchlorin derivative or ethylene glycol derivative or metal complex thereof, to reach and remain at the affected area, be absorbed transdermally, accumulate at the diseased area, and kill target cells or bacteria by ultrasonic irradiation. According to the inventors' studies, it has been found that a sufficient effect can be obtained if the amount is 0.1 to 20% by weight based on the weight of the preparation.
配合量が０．１重量％未満であると目的とする治療効果を上げることができず、また２０重量％以上配合させてもそれ以上の効果は得られなかった。
なお、配合量は含有させる有効成分の種類により一概に特定することはできず、また、含有させる有効成分の安定性は有効成分の濃度、用いる基剤に大きく影響されないため、上記の含有量の範囲内で、用途に合わせ種々変更させることが可能である。
If the amount was less than 0.1% by weight, the desired therapeutic effect could not be achieved, and even if the amount was more than 20% by weight, no further effect could be obtained.
The amount of the active ingredient to be added depends on the type of active ingredient contained, and cannot be generally determined. Furthermore, since the stability of the active ingredient contained is not significantly affected by the concentration of the active ingredient or the base used, the amount can be varied in various ways within the above content range according to the application.
以上のようにして得られた本発明の製剤をＳＤＴに使用する場合には、各種癌には点滴注射、舌下投与、坐剤投与又は患部位に局部注射することにより、他方、日光角化症、炎症性角化症、表皮癌、感染症等には皮膚疾患部位に直接塗布することにより、効果的にフェニルクロリン誘導体又はエチレングリコール誘導体あるいはその金属錯体が集積され、その後、その部位を超音波等の照射により、当該疾患を効果的に治療することができる。 When the preparation of the present invention obtained as described above is used for SDT, various cancers can be treated by intravenous injection, sublingual administration, suppository administration, or local injection into the affected area, while actinic keratosis, inflammatory keratosis, epidermal cancer, infectious diseases, etc. can be treated by direct application to the affected skin area, whereby the phenylchlorin derivative, ethylene glycol derivative, or metal complex thereof can be effectively accumulated, and then the affected area can be irradiated with ultrasound or the like to effectively treat the disease.
この軟膏剤、ローション剤等の外用剤の適用において、本発明が提供するフェニルクロリン誘導体又はエチレングリコール誘導体あるいはその金属錯体は、テープストリッピングを必要としないで皮膚患部への浸透性が良好なものであり、したがって、皮膚疾患治療におけるＳＤＴにおいて、患者に負担を与えることがなく、治療自体を簡便に行える利点を有している。
なお、軟膏剤、ローション剤等の外用剤の塗布にあたっては、ＯＤＴ効果（密封包帯効果：Occlusive Dressing Technique）を得るために、塗布部位を密閉状態に保つこともより効果的である。
In application of topical preparations such as ointments and lotions, the phenylchlorin derivatives or ethylene glycol derivatives or their metal complexes provided by the present invention have good permeability to affected areas of the skin without the need for tape stripping, and therefore have the advantage that in SDT for treating skin diseases, the treatment itself can be carried out easily without imposing a burden on the patient.
When applying topical preparations such as ointments and lotions, it is more effective to keep the application site sealed in order to obtain an ODT effect (Occlusive Dressing Technique).
また、本発明のフェニルクロリン誘導体又はエチレングリコール誘導体あるいはそれらの金属錯体は、ＳＤＴ用の抗菌剤として、各種の感染症に対して有効であることが判明した。そのような感染症としては、ヒトのみならず各種動物における感染症を含み、酵母様真菌に起因する犬、猫等におけるマラセチア性外耳炎、爪感染症、水虫等の真菌による感染症、ＭＲＳＡ、緑膿菌、大腸菌等を原因菌とする各種感染症、歯周病菌による歯槽膿漏等の歯科系感染症、その他結核菌、或いはウイルスによる感染症や疣へのＳＤＴ用の治療に使用することができる。 The phenylchlorin derivatives or ethylene glycol derivatives or metal complexes thereof of the present invention have also been found to be effective as antibacterial agents for SDT against various infectious diseases. Such infectious diseases include those in various animals as well as humans, and can be used for SDT treatment of fungal infections such as Malassezia otitis externa, nail infections, and athlete's foot in dogs and cats caused by yeast-like fungi, various infectious diseases caused by bacteria such as MRSA, Pseudomonas aeruginosa, and Escherichia coli, dental infections such as pyorrhea caused by periodontal disease bacteria, and other infections and warts caused by tuberculosis bacteria or viruses.
本発明が提供するフェニルクロリン誘導体又はエチレングリコール誘導体あるいはその金属錯体を含有する製剤を塗布した後の疾患部位における超音波照射に際しては、種々の超音波を使用することができる。なかでも、医療用の超音波照射器等を用いることがより効果的である。 Various types of ultrasound can be used to irradiate the diseased area after application of the preparation containing the phenylchlorin derivative or ethylene glycol derivative or its metal complex provided by the present invention. In particular, it is more effective to use a medical ultrasound irradiator.
かくして、本発明の注射製剤あるいは軟膏製剤、ローション製剤等を疾患部位に注射しあるいは塗布し、有効成分を患部に集積あるいは経皮吸収させた後、当該疾患部位を超音波照射することにより、当該癌疾患、皮膚疾患、或いは感染症を効果的に治療することができる。 Thus, the injectable formulation, ointment formulation, lotion formulation, etc. of the present invention is injected or applied to the diseased area, the active ingredient is accumulated in the affected area or absorbed percutaneously, and then the diseased area is irradiated with ultrasound, thereby making it possible to effectively treat the cancer disease, skin disease, or infectious disease.
以下に本発明を、具体的処方例、試験例等により詳細に説明するが、本発明はこれらのものに限定されるものではない。 The present invention will be described in detail below with specific formulation examples, test examples, etc., but the present invention is not limited to these.
試験例１：フェニルクロリン誘導体（Ｃｏｍｐｏｕｎｄ Ａ、Ｃｏｍｐｏｕｎｄ Ｂ、Ｃｏｍｐｏｕｎｄ Ｆ、及びＣｏｍｐｏｕｎｄ Ｇ）の光毒性試験
本発明のフェニルクロリン誘導体又はエチレングリコール誘導体あるいはその金属錯体は、光無感作が望まれる。そのため、光処理に対する光毒性を、ＬＥＤ６６０ｎｍおよび４０５ｎｍによる処理に対する光毒性で検討した。
なお、同時に光陽性対照群として前願物質のＴＯＮＳ５０４（特許文献３）を用いた。
Test Example 1: Phototoxicity test of phenylchlorin derivatives (Compound A, Compound B, Compound F, and Compound G) The phenylchlorin derivatives or ethylene glycol derivatives or their metal complexes of the present invention are desired to be photoinsensitive. Therefore, the phototoxicity against light treatment was examined by the phototoxicity against treatment with LED of 660 nm and 405 nm.
At the same time, the aforementioned substance TONS504 (Patent Document 3) was used as a light positive control group.
＜実験操作＞
（１）９６ウェルプレートに１×１０３ cells/wellずつマウスメラノーマ細胞（Ｂ１０－Ｆ１０）又はヒト繊維芽細胞（ＨＤＦ）を別々に播種し、５％炭酸ガスインキュベーターで培養した。
（２）各化合物を１００μＭになるようにＤＭＳＯで溶解し、終濃度１μＭになるように培地で希釈した。
（３）培地交換で化合物入り培地を添加し、５時間後に１×ＰＢＳ（リン酸緩衝生理食塩水）で２回洗浄した後ＰＢＳを入れ，以下の条件でＬＥＤ光照射を行った。
<Experimental Procedure>
(1) Mouse melanoma cells (B10-F10) or human fibroblast cells (HDF) were seeded separately at 1×10 3 cells/well on a 96-well plate and cultured in a 5% carbon dioxide incubator.
(2) Each compound was dissolved in DMSO to a concentration of 100 μM, and then diluted with medium to a final concentration of 1 μM.
(3) The medium containing the compound was added during medium replacement, and after 5 hours, the cells were washed twice with 1x PBS (phosphate buffered saline), and then PBS was added. LED light irradiation was performed under the following conditions.
＜照射条件＞
・波長：４０５ｎｍ又は６６０ｎｍ
・照射量：４６．１ｍＷ／ｃｍ２
・照射時間：６分
<Irradiation conditions>
Wavelength: 405 nm or 660 nm
・Irradiation amount: 46.1mW/ cm2
- Irradiation time: 6 minutes
（４）照射４８時間後に、１×ＰＢＳで２回洗浄した後、０．５％クリスタルバイオレット（Crystal violet）溶液を５０μＬ添加した。
（５）常温で１５分間インキュベート後、水道水１００μＬ／wellで４回洗浄を行った。
（６）常温で１時間乾燥後、１％ＳＤＳを１００μＬ／well添加し、Crystal violetが溶出したのを確認後、マイクロプレートリーダーで吸光度（abs：５７０ｎｍ）を測定し、生存率を求めた。
(4) 48 hours after irradiation, the cells were washed twice with 1×PBS, and then 50 μL of 0.5% crystal violet solution was added.
(5) After incubation at room temperature for 15 minutes, the plate was washed four times with 100 μL/well of tap water.
(6) After drying at room temperature for 1 hour, 100 μL/well of 1% SDS was added. After confirming that crystal violet had been eluted, the absorbance (abs: 570 nm) was measured using a microplate reader to calculate the survival rate.
＜結果＞
その結果を図１、図２および図３に示した。図中に示した結果からも判明するように、本発明のフェニルクロリン誘導体又はエチレングリコール誘導体およびそれらのＭｎ錯体は、Ｂ１０－Ｆ１０細胞およびＨＤＦ細胞を用いた光照射実験で安定であること、すなわち光毒性が無いことが確認された。
一方、対照群である前願物質のＴＯＮＳ５０４やＣｏｍｐｏｕｎｄ Ｆには、光感作が認められた。
<Results>
The results are shown in Figures 1, 2 and 3. As is apparent from the results shown in the figures, the phenylchlorin derivatives or ethylene glycol derivatives and their Mn complexes of the present invention were confirmed to be stable, i.e., to have no phototoxicity, in the light irradiation experiments using B10-F10 cells and HDF cells.
On the other hand, photosensitization was observed in the control groups, that is, the precursor substances TONS504 and Compound F.
試験例２：フェニルクロリン誘導体又はエチレングリコール誘導体およびそのＭｎ金属錯体（Ｃｏｍｐｏｕｎｄ Ａ、Ｃｏｍｐｏｕｎｄ Ｂ、Ｃｏｍｐｏｕｎｄ Ｆ、及びＣｏｍｐｏｕｎｄ Ｇ）の細胞毒性試験
本発明のフェニルクロリン誘導体又はエチレングリコール誘導体およびそのＭｎ金属錯体は、暗所下での細胞無毒性が望まれる。そのための確認を、以下の試験により行った。
Test Example 2: Cytotoxicity test of phenylchlorin derivatives or ethylene glycol derivatives and their Mn metal complexes (Compound A, Compound B, Compound F, and Compound G) It is desired that the phenylchlorin derivatives or ethylene glycol derivatives and their Mn metal complexes of the present invention are non-cytotoxic in the dark. This was confirmed by the following test.
＜実験操作＞
（１）９６ウェルプレートに２．５×１０３cells/wellずつＢ１０－Ｆ１０細胞およびＨＤＦ細胞を別々に播種し、５％炭酸ガスインキュベーターで培養した。
（２）Ｃｏｍｐｏｕｎｄ Ａ、Ｃｏｍｐｏｕｎｄ Ｂ、Ｃｏｍｐｏｕｎｄ Ｆ、及びＣｏｍｐｏｕｎｄ Ｇを測り取り、濃度が夫々１０ｍＭ、３０ｍＭ、１０ｍＭ、３ｍＭ、１ｍＭ、０．３ｍＭ、０．１ｍＭ、０．０３ｍＭ及び０．０１ｍＭになるようにＤＭＳＯで調整した。
（３）ＤＭＳＯが１％になるようにＤＥＭＥ培地を用いて希釈し、終濃度を１０００μＭ、３００μＭ、１００μＭ、３０μＭ、１０μＭ、３μＭ、１μＭ、０．３μＭ及び０．１μＭにし、培地交換で添加した。
（４）添加２４時間後に1×ＰＢＳで２回洗浄を行い、培地を入れインキュベートした。
（５）２４時間後、１×ＰＢＳで２回洗浄を行い、０．５％Crystal violet溶液を５０μＬ添加した。
（６）常温で１５分間インキュベート後、水道水１００μＬ／wellで４回洗浄を行った。
（７）常温で１時間乾燥後、１％ＳＤＳを１００μＬ／well添加し、Crystal violetが溶出したのを確認後、マイクロプレートリーダーで吸光度（abs：５７０ｎｍ）を測定し、生存率を求めた。
<Experimental Procedure>
(1) B10-F10 cells and HDF cells were separately seeded at 2.5×10 3 cells/well on a 96-well plate and cultured in a 5% carbon dioxide incubator.
(2) Compound A, Compound B, Compound F, and Compound G were measured out and adjusted with DMSO to concentrations of 10 mM, 30 mM, 10 mM, 3 mM, 1 mM, 0.3 mM, 0.1 mM, 0.03 mM, and 0.01 mM, respectively.
(3) DMSO was diluted with DMEM medium to 1% to give final concentrations of 1000 μM, 300 μM, 100 μM, 30 μM, 10 μM, 3 μM, 1 μM, 0.3 μM and 0.1 μM, and added when the medium was replaced.
(4) 24 hours after addition, the cells were washed twice with 1x PBS, and the medium was added and incubated.
(5) After 24 hours, the plate was washed twice with 1x PBS, and 50 μL of 0.5% crystal violet solution was added.
(6) After incubation at room temperature for 15 minutes, the plate was washed four times with 100 μL/well of tap water.
(7) After drying at room temperature for 1 hour, 100 μL/well of 1% SDS was added. After confirming that crystal violet had been eluted, the absorbance (abs: 570 nm) was measured using a microplate reader to calculate the survival rate.
＜結果＞
その結果を図４、図５、図６、図７、図８および図９に示した。図中に示した結果からも判明するように、本発明のフェニルクロリン誘導体又はエチレングリコール誘導体およびそれらのＭｎ金属錯体（Ｃｏｍｐｏｕｎｄ Ａ、Ｃｏｍｐｏｕｎｄ Ｂ、及びＣｏｍｐｏｕｎｄ Ｇ）には、細胞毒性が１０μＭ以下では認められなかった。
なお、Ｃｏｍｐｏｕｎｄ Ａに関しては、１００μＭ以上の濃度では少し沈殿が出るようであった。
<Results>
The results are shown in Figures 4, 5, 6, 7, 8 and 9. As is clear from the results shown in the figures, the phenylchlorin derivatives or ethylene glycol derivatives of the present invention and their Mn metal complexes (Compound A, Compound B and Compound G) had no cytotoxicity at 10 μM or less.
Regarding Compound A, a small amount of precipitation appeared at concentrations of 100 μM or more.
試験例３：フェニルクロリン誘導体及びエチレングリコール誘導体（Ｃｏｍｐｏｕｎｄ Ａ、Ｃｏｍｐｏｕｎｄ Ｂ、Ｃｏｍｐｏｕｎｄ Ｆ、及びＣｏｍｐｏｕｎｄ Ｇ）の超音波活性試験
本発明のフェニルクロリン誘導体又はエチレングリコール誘導体およびそのＭｎ金属錯体は、超音波感作が望まれる。そのため、超音波処理に対する感受性を，以下のとおり検討した。
Test Example 3: Ultrasonic activity test of phenylchlorin derivatives and ethylene glycol derivatives (Compound A, Compound B, Compound F, and Compound G) The phenylchlorin derivatives or ethylene glycol derivatives of the present invention and their Mn metal complexes are desired to be sensitized by ultrasound. Therefore, the sensitivity to ultrasound treatment was examined as follows.
＜実験操作＞
（１）Φ６シャーレに１×１０５cells/wellずつＢ１０－Ｆ１０細胞およびＨＤＦ細胞を別々に播種し、５％炭酸ガスインキュベーターで培養した。
（２）各化合物を１００μＭになるようにＤＭＳＯで溶解し、終濃度１μＭになるように培地で希釈した。
（３）培地交換で化合物入り培地を添加し、５時間後に以下の条件で、超音波照射を行った。
<Experimental Procedure>
(1) B10-F10 cells and HDF cells were separately seeded at 1×10 5 cells/well in a Φ6 petri dish and cultured in a 5% carbon dioxide incubator.
(2) Each compound was dissolved in DMSO to a concentration of 100 μM, and then diluted with medium to a final concentration of 1 μM.
(3) The medium containing the compound was added during medium replacement, and after 5 hours, ultrasonic irradiation was performed under the following conditions.
＜照射条件＞
・Probe L（有効照射面積：５ｃｍ２）
・照射時間：４分
・Duty cycle：２０％
・周波数：１ＭＨｚ
・照射量：０．４Ｗ／ｃｍ２
<Irradiation conditions>
・Probe L (effective irradiation area: 5 cm2 )
・Irradiation time: 4 minutes ・Duty cycle: 20%
Frequency: 1MHz
・Irradiation amount: 0.4W/ cm2
（４）照射２４時間後に１×ＰＢＳで２回洗浄し、1×Trypsinを５００μＬ加え３分間インキュベート後、物理的に細胞をはがし１ｍＬ培地を加えた。
（５）細胞懸濁液３０μＬと０．４％トリパンブルー（Trypan blue）３０μＬを合わせ、血球計算盤を用いて生細胞数をカウントし、生存率を求めた。
(4) 24 hours after irradiation, the cells were washed twice with 1× PBS, 500 μL of 1× Trypsin was added, and the cells were incubated for 3 minutes. Then, the cells were physically detached and 1 mL of medium was added.
(5) 30 μL of the cell suspension and 30 μL of 0.4% Trypan blue were combined, and the number of live cells was counted using a hemocytometer to calculate the survival rate.
＜結果＞
その結果を図１０、図１２に示した。図中に示した結果からも判明するように、本発明のフェニルクロリンクロリン誘導体又はエチレングリコール誘導体およびその金属錯体は、Ｂ１６－Ｆ１０細胞並びにＨＤＦ細胞を用いた超音波照射実験で、細胞破壊効果があることが確認された。
なお、図の中でＵＳは超音波照射を、ＡはＣｏｍｐｏｕｎｄ Ａを、Ａ＋ＵＳはＣｏｍｐｏｕｎｄ Ａに超音波照射を、ＢはＣｏｍｐｏｕｎｄ Ｂを、Ｂ＋ＵＳはＣｏｍｐｏｕｎｄ Ｂに超音波照射を、Ｆ＋ＵＳはＣｏｍｐｏｕｎｄ Ｆに超音波照射を、Ｇ＋ＵＳはＣｏｍｐｏｕｎｄ Ｇに超音波照射をした結果を示す。
<Results>
The results are shown in Figures 10 and 12. As is clear from the results shown in the figures, it was confirmed that the phenylchlorine chlororin derivatives or ethylene glycol derivatives and their metal complexes of the present invention have a cell destruction effect in an ultrasound irradiation experiment using B16-F10 cells and HDF cells.
In the figure, US indicates ultrasonic irradiation, A indicates Compound A, A+US indicates ultrasonic irradiation of Compound A, B indicates Compound B, B+US indicates ultrasonic irradiation of Compound B, F+US indicates ultrasonic irradiation of Compound F, and G+US indicates ultrasonic irradiation of Compound G.
以下に、本発明が提供する新規フェニルクロリン誘導体又はエチレングリコール誘導体およびその金属錯体を含有する製剤について、ＳＤＴによる治療効果の実際を記載する。 The actual therapeutic effect of SDT for the preparations containing the novel phenylchlorin derivatives or ethylene glycol derivatives and their metal complexes provided by the present invention will be described below.
試験例４：担癌動物を用いた超音波照射試験
＜方法＞
Ｂａｌｂ/ｃ系の６週令雌性マウスを用いた。
担癌マウスは、ＥＭＴ－６腫瘍細胞を１×１０７個を背中背部に接種して作成した。
接種６日後に、本発明のフェニルクロリン誘導体又はエチレングリコール誘導体およびその金属錯体（Ｃｏｍｐｏｕｎｄ Ｂ及びＣｏｍｐｏｕｎｄ Ｇ）を尾静脈より夫々１ｍｇ／ｋｇ、５ｍｇ／ｋｇ又は１０ｍｇ／ｋｇを投与し、投与４時間後に５分間の超音波照射を行った。
Test Example 4: Ultrasound irradiation test using tumor-bearing animals <Method>
Six-week-old female mice of the Balb/c strain were used.
Tumor-bearing mice were generated by inoculating 1×10 7 EMT-6 tumor cells into the dorsal region.
Six days after the inoculation, the phenylchlorin derivative or ethylene glycol derivative of the present invention and its metal complex (Compound B and Compound G) were administered at 1 mg/kg, 5 mg/kg or 10 mg/kg, respectively, via the tail vein, and 4 hours after administration, ultrasonic irradiation was performed for 5 minutes.
＜超音波照射条件＞
・伊藤超短波（株）製装置：ＵＳＴ７７０
・周波数：１ＭＨｚ
・照射量：２Ｗ／ｃｍ２
・Duty cycle：５０％
<Ultrasonic irradiation conditions>
- Ito Ultrasonics Co., Ltd.: UST770
Frequency: 1MHz
・Irradiation amount: 2W/ cm2
Duty cycle: 50%
＜結果＞
その結果を図１３および図１４に示した。図中に示した結果からも判明するように、本発明のフェニルクロリン誘導体又はエチレングリコール誘導体およびその金属錯体には腫瘍抑制効果が見られることが確認され、特に、Ｃｏｍｐｏｕｎｄ ＢおよびＣｏｍｐｏｕｎｄ Ｇを用いた超音波照射では、薬剤１ｍｇ、５ｍｇおよび１０ｍｇ投与後、４時間後に５分間の超音波照射で顕著な効果が認められた。
<Results>
The results are shown in Figures 13 and 14. As is clear from the results shown in the figures, it was confirmed that the phenylchlorin derivatives or ethylene glycol derivatives and their metal complexes of the present invention have a tumor suppressing effect, and in particular, in the case of ultrasonic irradiation using Compound B and Compound G, a remarkable effect was observed when ultrasonic irradiation was performed for 5
以上記載のように、本発明は超音波化学療法（ＳＤＴ）として使用する電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体およびそれらの金属錯体を提供するものであり、本発明が提供する電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体およびその金属錯体は、癌適用注射製又は剤及び舌下剤や皮膚適用軟膏製剤とすることにより、患部到達性や皮膚透過性（経皮吸収性）が極めて良好なものである。
したがって、癌疾患部位に選択的に集積され、他方、日光角化症、炎症性角化症、表皮癌、感染症等の皮膚疾患、例えば乳頭腫等の疾患部位に塗布することにより、疾患部位に効果的に電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体およびその金属錯体が集積され、超音波照射により効果的に癌治療や皮膚疾患治療を行うことができる。
As described above, the present invention provides phenylchlorin derivatives or ethylene glycol derivatives carrying an electron-withdrawing group and metal complexes thereof for use in ultrasonic chemotherapy (SDT). The phenylchlorin derivatives or ethylene glycol derivatives carrying an electron-withdrawing group and metal complexes thereof provided by the present invention have extremely good delivery to the affected area and skin permeability (transdermal absorbability) when made into injections or preparations for use in cancer, sublingual preparations, or ointments for use on the skin.
Therefore, the phenylchlorin derivatives or ethylene glycol derivatives and their metal complexes carrying an electron-withdrawing group are selectively accumulated at the site of cancer disease, and by applying the same to the site of a skin disease such as actinic keratosis, inflammatory keratosis, epidermal cancer, or an infectious disease, e.g., papilloma, the phenylchlorin derivatives or ethylene glycol derivatives and their metal complexes carrying an electron-withdrawing group are effectively accumulated at the site of the disease, and cancer or skin disease can be effectively treated by ultrasonic irradiation.
また、ＳＤＴ用の抗菌剤として、各種細菌に対して抗菌活性を示すものであり、今日の感染症治療用いられている抗生物質等は、薬剤耐性が容易に生じやすいものであるが、本発明のＳＤＴ診断・治療システムにおいては、そのような薬剤耐性の問題が発生しない点で、その利点は、極めて特異的なものであり、その医療上の価値は多大なものである。
また、過去の試験結果より，本電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体をＭｎ錯体化すると水溶性も増し、ＭＲＩ造影剤としても有効であり、本発明の電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体およびそのＭｎ金属錯体を投与後、患部をＭＲＩで特定後、ＳＤＴ照射治療が可能である利点を有する。
In addition, as an antibacterial agent for SDT, it exhibits antibacterial activity against various bacteria. Antibiotics currently used to treat infectious diseases are prone to develop drug resistance, but the SDT diagnosis and treatment system of the present invention has the advantage that such drug resistance does not occur, making it extremely unique and of great medical value.
Furthermore, past test results have shown that when the present electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative is complexed with Mn, its water solubility is increased and it is also effective as an MRI contrast agent, and has the advantage that after administration of the present electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative and its Mn metal complex, the affected area can be identified by MRI and then SDT irradiation treatment can be performed.
Claims (11)
Ｘは、次式：
Ｒは、－Ｎ(ＣＨ２ＣＯＯＮａ) ２［これはイミノ二酢酸の残基を示す］、アスパラギン酸残基或いはグルタミン酸残基、又は－ＮＨ(ＣＨ２ＣＨ２Ｏ）ｎＨ（ｎは、１から３の整数を表す）を表し、
Ｍは、Ｍｎ、Ｆｅ又はＣｕを表し、
Ｚは、配位子を示し、ＯＡｃ又はＣｌである］
で示される電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体の金属錯体又はその薬理学的に許容される塩。 The following formula (I):
X is of the following formula:
R represents -N(CH 2 COONa) 2 [which represents a residue of iminodiacetic acid], an aspartic acid residue, a glutamic acid residue, or -NH(CH 2 CH 2 O) n H (n represents an integer of 1 to 3);
M represents Mn, Fe or Cu;
Z represents a ligand and is OAc or Cl.
or a pharmacologically acceptable salt thereof.
で示される請求項１に記載の電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体の金属錯体又はその薬理学的に許容される塩。 Formula (I) is the following formula (I)-A type:
2. The metal complex of the electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative according to claim 1, which is represented by the following formula: or a pharmacologically acceptable salt thereof.
で示される請求項１に記載の電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体の金属錯体又はその薬理学的に許容される塩。 Formula (I) is of the following formula (I)-B type:
2. The metal complex of the electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative according to claim 1, which is represented by the following formula: or a pharmacologically acceptable salt thereof.
Ｘは、次式：
Ｒは、－Ｎ(ＣＨ２ＣＯＯＮａ) ２［これはイミノ二酢酸の残基を示す］、アスパラギン酸残基或いはグルタミン酸残基、又は－ＮＨ(ＣＨ２ＣＨ２Ｏ）ｎＨ（ｎは、１から３の整数を表す）を表す］
で示される電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体、又はその薬理学的に許容される塩。 The following formula (II):
X is of the following formula:
R represents --N(CH 2 COONa) 2 [which represents a residue of iminodiacetic acid], an aspartic acid residue, a glutamic acid residue, or --NH(CH 2 CH 2 O) n H (n represents an integer of 1 to 3)].
or a pharmacologically acceptable salt thereof.
請求項４に記載の電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体、又はその薬理学的に許容される塩
を有効成分として含有する、超音波化学療法（ＳＤＴ：Sonodynamic Therapy）用の癌疾患治療用剤。 A metal complex of the electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative according to any one of claims 1 to 3, or a pharmacologically acceptable salt thereof;
5. A cancer disease treatment agent for sonodynamic therapy ( SDT ), comprising the electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative, or a pharmacologically acceptable salt thereof, as defined in claim 4 as an active ingredient.
請求項４に記載の電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体、又はその薬理学的に許容される塩
を有効成分として含有する、超音波化学療法（ＳＤＴ：Sonodynamic Therapy）用の皮膚疾患治療用剤。 A metal complex of the electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative according to any one of claims 1 to 3, or a pharmacologically acceptable salt thereof;
A skin disease treatment agent for sonodynamic therapy ( SDT ), comprising the electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative, or a pharmacologically acceptable salt thereof, as defined in claim 4 as an active ingredient.
請求項４に記載の電子求引基担持フェニルクロリン誘導体又はエチレングリコール誘導体、又はその薬理学的に許容される塩
を有効成分として含有する、超音波化学療法（ＳＤＴ：Sonodynamic Therapy）用の抗菌剤。 A metal complex of the electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative according to any one of claims 1 to 3, or a pharmacologically acceptable salt thereof;
5. An antibacterial agent for sonodynamic therapy ( SDT ), comprising the electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative, or a pharmacologically acceptable salt thereof, as defined in claim 4 as an active ingredient.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004524372A (en) | 2001-01-23 | 2004-08-12 | ミラヴァント ファーマシューティカルズ インコーポレイテッド | Large-scale production of tetrapyrroles |
| JP3613599B2 (en) | 1996-10-01 | 2005-01-26 | ワイス株式会社 | Iminochlorine aspartic acid derivative |
| JP5179245B2 (en) | 2008-04-30 | 2013-04-10 | 功 阪田 | Skin disease treatment |
| JP5651426B2 (en) | 2010-10-25 | 2015-01-14 | 阪田 功 | Chlorine derivative |
| JP2018199649A (en) | 2017-05-29 | 2018-12-20 | 再生ファーマ株式会社 | Ultrasonic sensitizer |
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|---|---|---|---|---|
| JP3613599B2 (en) | 1996-10-01 | 2005-01-26 | ワイス株式会社 | Iminochlorine aspartic acid derivative |
| JP2004524372A (en) | 2001-01-23 | 2004-08-12 | ミラヴァント ファーマシューティカルズ インコーポレイテッド | Large-scale production of tetrapyrroles |
| JP5179245B2 (en) | 2008-04-30 | 2013-04-10 | 功 阪田 | Skin disease treatment |
| JP5651426B2 (en) | 2010-10-25 | 2015-01-14 | 阪田 功 | Chlorine derivative |
| JP2018199649A (en) | 2017-05-29 | 2018-12-20 | 再生ファーマ株式会社 | Ultrasonic sensitizer |
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| Anticancer Research，2011年，Vol.31，pp.501-506 |
| Cancer Science，2007年，Vol.98, No.6，pp.916-920 |
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